Recent strategies to reprogram cell fate have the potential to transform applications in disease modeling, drug screening, and gene and cell therapies for regenerative medicine. The ability to generate any cell type of interest from an easily-acquired starting cell holds great promise for the creation of next-generation models and therapies for personalized medicine. There has been a recent expansion in cell reprogramming methods following the discovery that somatic cells can be converted to a pluripotent state by overexpression of a cocktail of transcription factors. However, it still remains a pertinent challenge to generate cell types with functionally mature phenotypes at high efficiency. To address this bottleneck, we are developing new strategies to identify and relieve barriers to cell reprogramming. We propose to utilize next-generation epigenome editing tools based on the programmable CRISPR/Cas9 system to modulate the endogenous epigenome to facilitate reprogramming outcomes. In this project, we will use CRISPR/Cas9-based epigenetic modifiers to remodel endogenous chromatin to initiate the generation of induced neurons and elucidate regulatory mechanisms that govern neuronal fate specification. We will exploit the high-throughput capacity of the CRISPR/Cas9 system for (1) the unbiased identification of transcription factor combinations that maximize production of induced neurons and (2) the identification of essential regulatory elements that govern neuronal differentiation and phenotype. The insights gained from these studies will have broad relevance to improving cell reprogramming strategies and enhancing our understanding of cell differentiation and plasticity.